Laboratoire de Bactériologie-Virologie-Hygiène, Hôpital Henri Mondor, Assistance Publique-Hôpitaux de Paris, Créteil, France.
Ann Clin Microbiol Antimicrob. 2010 Aug 4;9:21. doi: 10.1186/1476-0711-9-21.
Pseudomonas aeruginosa is responsible for numerous bloodstream infections associated with severe adverse outcomes in case of inappropriate initial antimicrobial therapy. The present study was aimed to develop a novel quantitative PCR (qPCR) assay, using ecfX as the specific target gene, for the rapid and accurate identification of P. aeruginosa from positive blood cultures (BCs).
Over the period August 2008 to June 2009, 100 BC bottles positive for gram-negative bacilli were tested in order to evaluate performances of the qPCR technique with conventional methods as gold standard (i.e. culture and phenotypic identification).
Thirty-three strains of P. aeruginosa, 53 strains of Enterobactericaeae, nine strains of Stenotrophomonas maltophilia and two other gram-negative species were isolated while 3 BCs were polymicrobial including one mixture containing P. aeruginosa. All P. aeruginosa clinical isolates were detected by qPCR except a single strain in mixed culture. Performances of the qPCR technique were: specificity, 100%; positive predictive value, 100%; negative predictive value, 98.5%; and sensitivity, 97%.
This reliable technique may offer a rapid (<1.5 h) tool that would help clinicians to initiate an appropriate treatment earlier. Further investigations are needed to assess the clinical benefit of this novel strategy as compared to phenotypic methods.
铜绿假单胞菌是导致血流感染的原因之一,若初始抗菌治疗不当,会导致严重的不良后果。本研究旨在开发一种新的实时荧光定量 PCR(qPCR)检测方法,以 ecfX 作为特异性靶基因,快速、准确地从阳性血培养物(BC)中鉴定出铜绿假单胞菌。
在 2008 年 8 月至 2009 年 6 月期间,检测了 100 个革兰氏阴性杆菌阳性的 BC 瓶,以评估 qPCR 技术与传统方法(即培养和表型鉴定)作为金标准的性能。
共分离出 33 株铜绿假单胞菌、53 株肠杆菌科细菌、9 株嗜麦芽窄食单胞菌和另外两种革兰氏阴性菌,3 个 BC 呈混合感染,其中一个混合培养物中含有铜绿假单胞菌。除了一株混合培养的菌株外,所有的铜绿假单胞菌临床分离株均通过 qPCR 检测到。qPCR 技术的性能为:特异性 100%;阳性预测值 100%;阴性预测值 98.5%;灵敏度 97%。
这种可靠的技术可能提供一种快速(<1.5 小时)的工具,有助于临床医生更早地开始进行适当的治疗。需要进一步的研究来评估与表型方法相比,这种新策略的临床获益。